[Properties and biocompatibility of collagen scaffold modified by genipin cross-linked L-lysine].

Collagen (Coll), as the basic material of matrix scaffolds for cell growth, has been widely used in the field of tissue engineering and regenerative medicine. In this study, collagen protein was modified by L-lysine (Lys), and cross-linked by genipin (GN) to prepare the L-lysine-modified collagen (Lys-Coll-GN) scaffolds. Microstructure, pore size, porosity, stability and biocompatibility of Lys-Coll-GN scaffolds were observed. The results showed that the bond between L-lysine and collagen protein molecule was formed by generating amide linkage, and mouse embryo fibroblasts proliferation was not inhibited in the Lys-Coll-GN scaffolds. In the multiple comparisons of Coll-scaf- folds, Coll-GN scaffolds and Lys-Coll-GN scaffolds, Coll-scaffolds was the worst in mechanical characteristics while the highest in biodegradation rate. Compared to Coll-GN scaffolds, Lys-Coll-GN scaffolds had more fiber structure, higher interval porosity (P<0. 01). Although the tensile stress of Lys-Coll-GN scaffolds reduced significantly, its e- longation length extended when the scaffolds was fractured (P<0. 01). The percentage of Lys-Coll-GN scaffolds residual weight was lower than that of Coll-GN scaffolds after all the scaffolds were treated by collagenase for 5 days (P<0. 01). This study suggested that Lys-Coll-GN scaffold had good biocompatibility, and it improved the mechanical property and degradation velocity for collagen-based scaffold. This study gave a new predominant type of tissue engineering scaffold for the regenerative medicine.

Injectable silk nanofiber hydrogels as stem cell carriers to accelerate wound healing.

Stem cells have potential utility in wound therapy, however the benefits are often limited due to cell injury from shear stress during injection and poor retention at the wound site. Here, shear-thinning silk nanofiber hydrogels were used to load bone marrow derived mesenchymal stem cells (BMSCs) and inject into wound sites to optimize cell retention and accelerate wound healing. The BMSCs in the silk nanofiber hydrogels maintained stemness better than the cells cultured on plates, and the expression of wound healing-related genes was significantly higher in the hydrogels with higher silk concentrations (2 wt%). The silk nanofibers physically prevented migration of BMSCs from the deposition site in the wound bed. In addition to faster wound healing, these BMSC-loaded hydrogels mediated angiogenesis and inflammation and improved collagen deposition and hair follicle regeneration in vivo in rats. Considering that these silk nanofiber hydrogels were successfully used here as carriers for stem cells to accelerate wound healing, further study for skin regeneration may be warranted.

A novel in situ-formed hydrogel wound dressing by the photocross-linking of a chitosan derivative.

In situ photopolymerized hydrogel dressings create minimally invasive methods that offer advantages over the use of preformed dressings such as conformability in any wound bed, convenience of application, and improved patient compliance and comfort. Here, we report an in situ-formed hydrogel membrane through ultraviolet cross-linking of a photocross-linkable azidobenzoic hydroxypropyl chitosan aqueous solution. The hydrogel membrane is stable, flexible, and transparent, with a bulk network structure of smoothness, integrity, and density. Fluid uptake ability, water vapor transmission rate, water retention, and bioadhesion of the thus resulted hydrogel membranes (0.1 mm thick) were determined to range from 97.0-96.3%, 2,934-2,561 g/m(2)/day, 36.69-22.94% (after 6 days), and 4.8-12.3 N/cm(2), respectively. These data indicate that the hydrogel membrane can maintain a long period of moist environment over the wound bed for enhancing reepithelialization. Specifically, these properties of the hydrogel membrane were controllable to some extent, by adjusting the substitution degree of the photoreactive azide groups. The hydrogel membrane also exhibited barrier function, as it was impermeable to bacteria but permeable to oxygen. In vitro experiments using two major skin cell types (dermal fibroblast and epidermal keratinocyte) revealed the hydrogel membrane have neither cytotoxicity nor an effect on cell proliferation. Taken together, the in situ photocross-linked azidobenzoic hydroxypropyl chitosan hydrogel membrane has a great potential in the management of wound healing and skin burn.

Water-insoluble amorphous silk fibroin scaffolds from aqueous solutions.

Regenerated silk fibroin (RSF) is emerging as promising biomaterial for regeneration, drug delivery and optical devices, with continued demand for mild, all-aqueous processes to control microstructure and the performance. Here, temperature control of assembly kinetics was introduced to prepare the water-insoluble scaffolds from neutral aqueous solutions of RSF protein. Higher temperatures were used to accelerate the assembly rate of the silk fibroin protein chains in aqueous solution and during the lyophilization process, resulting in water-insoluble scaffold formation. The scaffolds were mainly composed of amorphous states of the silk fibroin chains, endowing softer mechanical properties. These scaffolds also showed nanofibrous structures, improved cell proliferation in vitro and enhanced neovascularization and tissue regeneration in vivo than previously reported silk fibroin scaffolds. These results suggest utility of silk scaffolds in soft tissue regeneration.

Microskin-Inspired Injectable MSC-Laden Hydrogels for Scarless Wound Healing with Hair Follicles.

Scarless skin regeneration with functional tissue remains a challenge for full-thickness wounds. Here, mesenchymal stem cell (MSC)-laden hydrogels are developed for scarless wound healing with hair follicles. Microgels composed of aligned silk nanofibers are used to load MSCs to modulate the paracrine. MSC-laden microgels are dispersed into injectable silk nanofiber hydrogels, forming composites biomaterials containing the cells. The injectable hydrogels protect and stabilize the MSCs in the wounds. The synergistic action of silk-based composite hydrogels and MSCs stimulated angiogenesis and M1-M2 phenotype switching of macrophages, provides a suitable niche for functional recovery of wounds. Compared to skin defects treated with MSC-free hydrogels, the defects treated with the MSC-laden composite hydrogels heal faster and form scarless tissues with hair follicles. Wound healing can be further improved by adjusting the ratio of silk nanofibers and particles and the loaded MSCs, suggesting tunability of the system. To the best of current knowledge, this is the first time scarless skin regeneration with hair follicles based on silk material systems is reported. The improved wound healing capacity of the systems suggests future in vivo studies to compare to other biomaterial systems related to clinical goals in skin regeneration in the absence of scarring.

Biomineralization of stable and monodisperse vaterite microspheres using silk nanoparticles.

The influence of silk fibroin (SF) on calcium carbonate (CaCO3) biomineralization has been investigated; however, the formation of small, uniform SF-regulated vaterite microspheres has not been reported. In this work, spherical CaCO3 was synthesized via coprecipitation in the presence of SF. SF nanostructures were first tuned by self-assembly at 60 °C to provide better control of the nucleation of CaCO3. Subsequently, monodisperse vaterite microspheres about 1.1 µm were generated by controlling aggregation and growth of CaCO3 under appropriate concentrations of SF and Ca ions. In contrast to unstable vaterite, the microspheres generated in the present study have sufficient stability in aqueous solution for at least 8 days without transformation into calcite, due to the electrostatic interactions between the Ca ions and the preassembled SF nanostructures. The microspheres as drug carriers of doxorubicin (DOX) were assessed and found to have good encapsulation efficiency, sustained drug release without burst release, and pH sensitivity. These new SF/CaCO3 hybrids may provide new options for various biomedical applications.

The preparation, characterization and evaluation of regenerated cellulose/collagen composite hydrogel films.

Porous structured regenerated cellulose films were oxidized by periodate oxidation to obtain 2,3-dialdehyde cellulose (DARC) films, which were then reacted with collagen to obtain DARC/Col composite films. The subsequent FT-IR spectra indicated that collagen was immobilized on the DARC matrix via the Schiff base reaction between NH2 in collagen and CHO in DARC backbone. Scanning electron microscopy revealed that DARC/Col exhibited a refined 3D network structure and its porosity and pore size decreased with increasing of collagen concentration. The composite films demonstrated a good equilibrium-swelling ratio, air permeability and water retention properties. The composite films also showed excellent mechanical properties, which was vital for practical application. Finally, the cytotoxicity of the composite film was evaluated using NIH3T3 mice fibroblast cells, the results revealed that DARC/Col composite films have good biocompatibility for use as scaffold material in tissue engineering.